Huw Lee L, Hwa Lee K
Department of Veterinary Medicine, National Chung Hsing University, Taiwan, R.O.C.
J Virol Methods. 1997 Jan;63(1-2):113-9. doi: 10.1016/s0166-0934(96)02119-2.
The polymerase chain reaction (PCR) was used to amplify a 578-bp fragment of the fowl poxvirus (FPV) genome and with a set of primers framed a region within the gene coding for 4b core protein. An amplified product was detected with six strains of FPV, whereas none was obtained from uninfected cell cultures, skin tissue or four unrelated avian pathogens. The sensitivity of PCR was tested with nucleic acids from the FPV-infected cell cultures. The detection limit was 10(-1) TCID50 in an ethidium bromide-stained gel. In addition, this assay system was used to detect FPV in tissue specimens of skin and respiratory swabs collected from commercially reared chickens. The identity of the amplification products from the tissue specimen preparations was determined further by using a simple, rapid procedure in which an internally nested, end-labeled probe was used.
采用聚合酶链反应（PCR）扩增禽痘病毒（FPV）基因组的一个578 bp片段，所用引物框定了编码4b核心蛋白的基因内的一个区域。6株FPV均检测到扩增产物，而未感染的细胞培养物、皮肤组织或4种无关禽病原体均未获得扩增产物。用感染FPV的细胞培养物中的核酸检测PCR的灵敏度。在溴化乙锭染色凝胶中，检测限为10(-1) TCID50。此外，该检测系统用于检测从商业饲养鸡采集的皮肤组织标本和呼吸道拭子中的FPV。通过使用一种简单、快速的方法进一步确定组织标本制备物中扩增产物的身份，该方法使用内部嵌套的末端标记探针。
